AffiAB Goat Anti-mRuby Polyclonal Antibody for detection of mRuby-tagged proteins in immunofluorescence and cell imaging

Anti-mRuby Antibody

by Agennix Worldwide
Fluorescent Protein Antibody Guide

Detection of mRuby-Tagged Proteins in Cell Imaging and Localization Studies

Anti-mRuby is a research tool used to detect mRuby, a monomeric red fluorescent protein (RFP), and mRuby-tagged fusion proteins. It is applied in immunofluorescence (IF), immunohistochemistry (IHC), protein localization studies, and cell biology research workflows.

Primary target mRuby fluorescent protein and mRuby-tagged fusion proteins expressed in cells or tissues.
Key applications Immunofluorescence (IF), immunohistochemistry (IHC), subcellular localization, and tagged protein detection.
Research context Cell imaging, protein trafficking, expression confirmation, and multi-color fluorescence microscopy.

What Is an Antibody Anti-mRuby ?

An antibody anti-mRuby is a research-grade primary antibody that specifically recognizes and binds to the mRuby fluorescent protein. mRuby is a monomeric red fluorescent protein (mRFP) derived from the Entacmaea quadricolor sea anemone lineage. It is widely used as a genetically encoded fusion tag to visualize proteins of interest in living and fixed biological systems.

When a target protein is expressed as an mRuby fusion construct, an antibody anti-mRuby enables antibody-based detection of that tagged protein. This approach is particularly valuable when the intrinsic fluorescence of mRuby is insufficient after fixation, when signal amplification is needed, or when the experiment requires combining fluorescent protein detection with other immunostaining markers.

antibody anti-mRuby supports researchers in confirming mRuby-tagged protein expression, evaluating subcellular localization, and integrating mRuby detection into multi-channel imaging or co-staining workflows.

AffiAB® Goat Anti-mRuby Polyclonal Antibody

A polyclonal primary antibody raised in goat, designed for detection of mRuby fluorescent protein and mRuby-tagged fusion proteins in immunofluorescence, immunohistochemistry, and protein localization research.

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Why Use an antibody anti-mRuby?

Although mRuby is intrinsically fluorescent, antibody-based detection offers several practical advantages in research workflows. Chemical fixation can reduce or abolish the native fluorescence of mRuby, making antibody staining essential for fixed-sample analysis. Additionally, primary antibody detection followed by a fluorophore-conjugated secondary antibody provides signal amplification, improving sensitivity in samples with low mRuby expression levels.

Subcellular Protein Localization

Detect mRuby-tagged proteins within fixed cells or tissue sections to map subcellular distribution, organelle targeting, and localization dynamics.

Expression Confirmation

Verify successful expression of mRuby-tagged fusion constructs in transfected or transduced cell lines using antibody-based detection methods.

Fixed-Cell Immunofluorescence

Enable immunofluorescence staining of fixed and permeabilized samples where native mRuby fluorescence may be reduced or lost after chemical fixation.

Multi-Color Co-Staining

Combine antibody anti-mRuby detection with other primary antibodies to study co-localization, protein interactions, and spatial relationships within the same sample.

Common Applications of antibody anti-mRuby

Application How antibody anti-mRuby Is Used
Immunofluorescence (IF) Detects mRuby-tagged proteins in fixed cells; supports co-localization studies with other fluorescent markers or antibody targets.
Immunohistochemistry (IHC) Visualizes mRuby-tagged targets in formalin-fixed or frozen tissue sections when compatible antigen retrieval and detection systems are used.
Subcellular localization studies Maps the intracellular distribution of mRuby-tagged proteins across compartments such as the nucleus, cytoplasm, mitochondria, or plasma membrane.
Protein–protein interaction workflows Supports co-immunoprecipitation (co-IP) or proximity ligation assay (PLA) strategies involving mRuby-tagged bait proteins, depending on protocol compatibility.
Cell biology and trafficking research Enables study of protein trafficking, turnover, secretory pathway dynamics, and expression patterns of mRuby-tagged constructs in fixed samples.

Antibody anti-mRuby in Fluorescent Protein Research

Genetically encoded fluorescent protein tags — including GFP, mCherry, mRuby, mScarlet, and TagRFP — are foundational tools in modern cell biology. mRuby is a monomeric red fluorescent protein with excitation and emission peaks suitable for use alongside green and far-red fluorescent markers, making it well-suited for multi-color imaging experiments.

In these workflows, an antibody anti-mRuby provides an orthogonal detection strategy. Rather than relying solely on the intrinsic fluorescence of the mRuby tag, researchers can use antibody staining to amplify signal, confirm protein identity, or integrate mRuby detection into immunostaining panels that include antibodies against endogenous proteins.

This approach is especially relevant in fixed-tissue studies, high-content imaging, and experiments where signal-to-noise ratio is critical for accurate interpretation.

Research note: Staining quality depends on multiple experimental variables including fixation method, permeabilization conditions, blocking buffer composition, primary antibody dilution, secondary antibody selection, microscope filter configuration, and the expression level of the mRuby-tagged construct. Optimization is recommended for each new sample type or experimental system.

General Immunofluorescence Protocol Using antibody anti-mRuby

The following workflow outlines a standard immunofluorescence procedure for detecting mRuby-tagged proteins in fixed cells. All steps should be optimized for the specific cell line, fixation method, antibody concentration, and imaging system used.

1
Culture cells expressing the mRuby-tagged construct Grow transfected or transduced cells under appropriate conditions until they reach the desired confluency and expression level for imaging.
2
Wash and fix the sample Rinse cells with phosphate-buffered saline (PBS) and fix using a compatible fixative. A common choice is 4% paraformaldehyde (PFA) in PBS for 10–20 minutes at room temperature.
3
Permeabilize and block For intracellular targets, permeabilize cells using a detergent such as 0.1–0.3% Triton X-100 in PBS. Block non-specific binding with 1–5% BSA or normal serum in PBS for 30–60 minutes.
4
Incubate with anti-mRuby primary antibody Dilute the antibody anti-mRuby in blocking buffer at the recommended concentration and incubate with the sample, typically overnight at 4°C or for 1–2 hours at room temperature.
5
Apply a compatible secondary antibody After washing, incubate with a species-matched, fluorophore-conjugated secondary antibody. Protect from light during and after this step to preserve fluorophore integrity.
6
Counterstain, mount, and image Optionally add a nuclear counterstain such as DAPI or Hoechst. Mount the sample with an appropriate mounting medium and image using a fluorescence microscope with filters matched to the secondary antibody fluorophore.

Recommended Materials for Anti-mRuby Immunofluorescence

  • Cells or tissue samples expressing an mRuby-tagged protein of interest
  • Anti-mRuby primary antibody (e.g., AffiAB® Goat Anti-mRuby Polyclonal Antibody)
  • Species-matched, fluorophore-conjugated secondary antibody
  • Phosphate-buffered saline (PBS), pH 7.4
  • Fixative solution (e.g., 4% paraformaldehyde in PBS)
  • Permeabilization buffer (e.g., 0.1–0.3% Triton X-100 in PBS)
  • Blocking buffer (e.g., 1–5% BSA or normal serum in PBS)
  • Mounting medium with or without DAPI
  • Fluorescence microscope with appropriate excitation and emission filter sets

Tips for Optimizing antibody anti-mRuby Staining

Titrate the primary antibody

Begin with the supplier's recommended dilution range and perform a titration series to identify the concentration that gives the best signal-to-background ratio for your sample.

Include appropriate controls

Run non-transfected cells, no-primary-antibody controls, and known mRuby-positive samples in parallel to assess specificity, background, and staining consistency.

Protect fluorophores from light

Keep secondary antibody solutions and stained samples protected from light at all times to prevent photobleaching and preserve fluorescence signal quality.

Verify spectral compatibility

Choose secondary antibody fluorophores with minimal spectral overlap with other dyes or fluorescent proteins used in the same experiment to avoid bleed-through artifacts.

FAQ: Antibody anti-mRuby

What is an antibody anti-mRuby used for?
An antibody anti-mRuby is used to detect mRuby fluorescent protein and mRuby-tagged fusion proteins in research applications. Common uses include immunofluorescence (IF), immunohistochemistry (IHC), subcellular protein localization studies, and expression confirmation of mRuby-tagged constructs in cell and tissue samples.
Can antibody anti-mRuby be used for live-cell imaging?
Live-cell imaging typically relies on the intrinsic fluorescence of the mRuby tag without requiring an antibody. Antibody anti-mRuby is primarily used in fixed-cell or fixed-tissue workflows, where antibody-based signal amplification, co-staining with other markers, or detection after fixation-induced fluorescence loss is required.
Why use an antibody if mRuby is already fluorescent?
Chemical fixation can reduce or eliminate the native fluorescence of mRuby. Antibody-based detection restores the ability to detect the tagged protein in fixed samples, provides signal amplification for low-expression targets, and allows mRuby detection to be combined with other immunostaining markers in multi-channel experiments.
Which secondary antibody should be used with antibody anti-mRuby?
The secondary antibody must match the host species of the anti-mRuby primary antibody. For example, if the primary antibody is raised in goat, a species-matched anti-goat secondary antibody is required. The secondary antibody should be conjugated to a fluorophore compatible with the microscope filter set and spectrally distinct from other dyes used in the experiment.
Is antibody anti-mRuby for research use only?
Yes. antibody anti-mRuby is intended for research use only (RUO) and is not approved for diagnostic, therapeutic, or clinical applications. It should be used in accordance with the supplier's product documentation and validated laboratory protocols.
What is mRuby and how does it differ from mCherry or other red fluorescent proteins?
mRuby is a monomeric red fluorescent protein engineered for use as a genetically encoded tag in cell biology research. Like mCherry, it belongs to the red fluorescent protein (RFP) family, but mRuby has distinct spectral properties, brightness, and photostability characteristics. Researchers select between mRuby, mCherry, mScarlet, and other RFPs based on the specific requirements of their imaging system and experimental design.

Explore Antibody Anti-mRubyfor Tagged Protein Detection

AffiAB® Goat Anti-mRuby Polyclonal Antibody is a research-grade primary antibody for detection of mRuby and mRuby-tagged fusion proteins. It supports immunofluorescence, immunohistochemistry, subcellular localization studies, and multi-color fluorescence microscopy workflows.

View Antibody Anti-mRuby
by Agennix Worldwide

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